Pancreatic adenocarcinoma (PDA) is a lethal malignancy with a particularly poor prognosis due to the advanced nature of the disease at the time of diagnosis in most patients. Currently, there are no effective screening strategies for earl detection of PDA. Studies from our lab have demonstrated that pancreatic epithelial cells are shed into the blood stream at early stages of disease in genetically engineered mouse models as well as in patients with pre-malignant cystic lesions of the pancreas. These observations present a unique opportunity to use these cells as the basis for screening for PDA. The primary goal of this project is to develop a robust non-invasive test that can be used to detect early stage pancreatic ductal adenocarcinoma (PDA) based on the presence of circulating epithelial cells of pancreatic origin in blood. We will use magnetic nanoparticle technology and a microfluidic platform to enrich circulating epithelial cells (CECs) from whole blood followed by RNA-based detection, including RNA fluorescent in situ hybridization (RNA FISH) and quantitative PCR. We will validate this using a lineage-labeled genetically engineered mouse model of PDA in which CECs can be easily detected in the blood. Further, we will utilize the optimized device along with additional single cell isolation techniques such as the DEPArray(tm) technology or flow cytometry to perform next-generation sequencing on CECs in order to (i) identify novel biomarkers for more effective screening and diagnosis and (ii) investigate functional differences between CECs at different stages of pancreatic cancer progression. Finally we will implement a pilot study in a cohort of patients with known PDA to enrich and detect CECs in the blood using the assay developed in previous aims. We envision that this strategy could be adapted for a number of other applications including evaluating response to therapy, disease-staging and screening of high-risk populations, thereby leading to improvements in pancreatic cancer survival.